“Four-in-One” multifunctional nanohybrids integrated magnetic colorimetric catalytic SERS-driven lateral flow immunoassay for ultrasensitive detection of MPXV

A “Four-in-One” multifunctional nanotags-based TLFA is developed for ultrasensitive qualitative and quantitative detection of MPXV. The MS@Pt tags provide rapid magnetic enrichment of analytes in complex samples, avoiding complex pre-processing. The MS@Pt tags synergize three output signals: colorimetric, catalytically enhanced colorimetric, and SERS on LFA strips, increasing detection sensitivity and accuracy. • Au/Pt co-decorated Fe 3 O 4 (MS@Pt) NPs integrate four excellent properties in one. • Multifunctional MS@Pt NPs was used to construct triple-signal mode LFA (TLFA). • The MS@Pt-TLFA platform displays outstanding performance in MPXV detection. • The TLFA allows flexible switching of detection modes for different scenarios. The global outbreak of monkeypox in 2022 challenged the means of early, rapid, and accurate screening of the disease to avoid widespread outbreaks. However, as the most easy and convenient point-of-care testing (POCT) method, lateral flow immunoassay (LFA) is limited by the insufficient sensitivity and low reliability of visualizing single-signal patterns. Here, a triple-signal mode LFA (TLFA) using “four-in-one” multifunctional Au/Pt co-decorated Fe 3 O 4 (MS@Pt) nanotags is reported. The MS@Pt-DTNB tags with excellent magnetic, optical, peroxidase-like, and SERS properties outputs a triple-signal readout of colorimetric, catalytically enhanced colorimetric, and SERS signals for Monkeypox virus (MPXV) detection after magnetic separation. The qualitative and quantitative limits of detection (LODs) of MS@Pt-TLFA strips were 0.005 and 0.0016 ng mL −1 for MPXV A29L protein, respectively, and 73.78 and 17.724 pfu mL −1 for inactivated virus, respectively, which is 2–3 orders of magnitude more sensitive compared with the AuNP-based LFA. The diversity, complementarity, and mutual corroboration of integrated output signals in this MS@Pt-TLFA improve the flexibility, accuracy, and practicability of MPXV detection, providing a potential avenue for immunodiagnostic in a variety of application scenarios.